Simultaneous imaging and flow-cytometry-based detection of multiple fluorescent senescence markers in therapy-induced senescent cancer cells

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Item Type:	Article
Title:	Simultaneous imaging and flow-cytometry-based detection of multiple fluorescent senescence markers in therapy-induced senescent cancer cells
Creators Name:	Dovjak, E., Mairhofer, M., Wöß, C., Qi, J., Fan, D.N.Y., Schmitt, C.A. and Yu, Y.
Abstract:	Chemotherapeutic drugs can induce irreparable DNA damage in cancer cells, leading to apoptosis or premature senescence. Unlike apoptotic cell death, senescence is a fundamentally different machinery restraining propagation of cancer cells. Decades of scientific studies have revealed the complex pathological effects of senescent cancer cells in tumors and microenvironments that modulate cancer cells and stromal cells. New evidence suggests that senescence is a potent prognostic factor during cancer treatment, and therefore rapid and accurate detection of senescent cells in cancer samples is essential. This paper presents a method to visualize and detect therapy-induced senescence (TIS) in cancer cells. Diffuse large B-cell lymphoma (DLBCL) cell lines were treated with mafosfamide (MAF) or daunorubicin (DN) and examined for the senescence marker, senescence-associated β-galactosidase (SA-β-gal), the DNA synthesis marker 5-ethynyl-2′-deoxyuridine (EdU), and the DNA damage marker gamma-H2AX (γH2AX). Flow cytometer imaging can help generate high-resolution single-cell images in a short period of time to simultaneously visualize and quantify the three markers in cancer cells.
Keywords:	beta-Galactosidase, Biomarkers, Cellular Senescence, DNA Damage, Flow Cytometry, Neoplasms, Substances, Tumor Microenvironment
Source:	Journal of Visualized Experiments
ISSN:	1940-087X
Publisher:	JoVE
Number:	185
Page Range:	e63973
Date:	12 July 2022
Official Publication:	https://doi.org/10.3791/63973
PubMed:	View item in PubMed

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